Due to the rapidly increasing number of therapeutic humanized monoclonal antibodies being developed and FDA-approved, there is a clear need for improved cell line development to support recombinant therapeutic protein manufacturing. There is a current and projected shortage of worldwide GMP manufacturing capacity for these therapeutic proteins, and approaches for getting more protein from the available bioreactor capacity are deemed critical. Generation of stable high-secreting producer cell lines is a major bottleneck, requiring a 4-10 month process with no guarantee of success. Cloning of cells for high expression of a secreted product (e.g., antibody) is a particular challenge because the secreted product does not remain associated with the cell. The novel laser-enabled analysis and processing (LEAP(tm)) technology platform for high-throughput cell imaging and in situ laser-mediated cell manipulation has the potential to address these needs. Preliminary results shown within indicate significantly improved specific antibody secretion rates from LEAP-derived clones, along with an acceleration of the process, as compared to current methods. The goal of Phase I is to confirm these preliminary results and develop improvements by pursuing the following specific aims: (1) optimize in situ detection of secreted antibody and laser-mediated cell purification; (2) develop automation of cloning processes and monitoring of clone outgrowth; and (3) demonstrate impact of LEAP on cell cloning for recombinant humanized antibody production. Phase I success will lead to a Phase II program to improve, apply and extend the technology, leading to Phase III commercialization with broad impact on recombinant antibody production. In biopharmaceutical manufacturing, over $30 billion worth of therapeutic proteins are produced annually from large-scale culture of recombinant cell lines producing a secreted protein. The number of biopharmaceuticals in clinical development and being FDA approved each year is growing dramatically, resulting in a worldwide shortage of GMP manufacturing capacity for these proteins. Therefore, methods to improve the yield of protein within the current bioreactor capacity constraints are highly desirable. [unreadable] [unreadable]